Transplantation of human amniotic epithelial cells improves hindlimb function in rats with spinal cord injury

Background Human amniotic epithelial cells (HAECs), which have several characteristics similar to stem cells, therefore could possibly be used in cell therapy without creating legal or ethical problems. In this study, we transplanted HEACs into the injured spinal cord of rats to investigate if the cells can improve the rats’ hindlimb motor function. Methods HAECs were obtained from a piece of fresh amnion, labeled with Hoechst33342, and transplanted into the site of complete midthoracic spinal transections in adult rats. The rats (n=21) were randomly divided into three groups: Sham-operation group (n=7), cells-graft group (n=7), and PBS group (n=7). One rat of each group was killed for histological analysis at the second week after the transplantation. The other six rats of each group were killed for histological analysis after an 8-week behavioral testing. Hindlimb motor function was assessed by using the open-field BBB scoring system. Survival rate of the graft cells was observed at second and eighth weeks after the transplantation. We also detected the myelin sheath fibers around the lesions and the size of the axotomized red nucleus. A one-way ANOVA was used to compare the means among the groups. The significance level was set at P<0.05.Results The graft HAECs survived for a long time (8 weeks) and integrated into the host spinal cord without immune rejection. Compared with the control group, HAECs can promote the regeneration and sprouting of the axons, improve the hindlimb motor function of the rats (BBB score: cells-graft group 9.0±0.89 vs PBS group 3.7±1.03, P<0.01), and inhibit the atrophy of axotomized red nucleus [cells-graft group (526.47±148.42) µm(2 )vs PBS group (473.69±164.73) µm(2), P<0.01]. Conclusion Transplantation of HAECs can improve the hindlimb motor function of rats with spinal cord injury.     

Brain-derived neurotrophical factor after olfactory ensheathing cells transplantation in spinal cord injury of rats

<正>Objective To observe the expression of brain - derived neurotrophical factor ( BDNF) in injury spinal cord after transplantation olfactory ensheathing cells ( OECs) , and to investigate the mechanism of OECs repairing spinal cord injury. Methods OECs from GFP transgenic rats were separated and cultured for transplantation. Spinal cord injury rats were separated two groups by random     

Antioxidation of melatonin against spinal cord injury in rats

Background The iron catalyzed lipid peroxidation plays an important role in the autodestruction of the injured spinal cord. This study was to detect the antioxidation of melatonin against spinal cord injury(SCI) in rats.Methods Sity Sprague-Dawley rats were randomly divided into four groups: group A (n=15) for laminectomyanly, group B (n=15) for laminectomy with SCI, group C (n = 15) for SCI and intraperitoneal injection of a bolus of 100 mg/kg melatonin, and group D (n = 15 ) for SCI and intraperitoneal injection of saline containing 5% ethanol. The SCI of animal model was made using modified Allen‘s method on T12. Six rats of each group were sacrificed 4 hours after injury, and the levels of free iron and malondialdehyde (MDA) of the involved spinal cord segments were measured by the bleomycin assay and thiobarbituric acid (TBA) separately. Functional recovery of the spinal cord was assessed by Modified Tarlov‘s scale and the inclined plane method at 1, 3, 7, 14, 21 days after SCI. The histologic changes of the damaged spinal cord were also examined at 7 days after SCI.Results After SCI, the levels of free iron and MDA were increased significantly and the modified Tarlov‘s score and inclined plane angle decreased significantly in groups B and D. In group C, the Tarlov‘s score and inclined plane angle were increased significantly at 7, 14 and 21 days, with histological improvement.Conclusion: Melatonin can reduce the level of lipid peroxidation and prevent damage to the spinal cord of rat.     

促红细胞生成素通过降低TSP-1和TGF-β表达对脊髓损伤保护作用的研究

Background Erythropoietin (EPO) functions as a tissue-protective cytokine in addition to its crucial hormonal role in red cell production and neuron protection. This study was designed to determine the erythropoietin neuron protection on experimental spinal cord trauma injury (SCI) rats by exploring the level of thrombospondin-1(TSP-1) and transforming growth factor-β (TGF-β) in the development of rats’ spinal cord injury (SCI) model. Methods Sixty Sprague–Dawley (SD) rats were divided into three groups at random (the sham operation control group, SCI group and erythropoietin treatment group). Using a weight-drop contusion SCI model in SCI group and erythropoietin treatment group, then rats were sacrificed at 24 h and 7d. BBB scores were checked as locomotor function. Pathological changes were also explored by H-E staining. The expression of TSP-1 and TGF-β were determined by immunohistochemistry staining analysis and Western Blot analysis respectively. Results Slighter locomotor dysfunction and more quickly recovered as their higher BBB scores were found in erythropoietin treatment group than that in SCI group (P < 0.01). Pathological observation also showed progressive disruption of the dorsal white and few neurons regeneration in SCI rats. TSP-1 and TGF-β expression increased at 24 h and 7 d after SCI in the injured segment. We observed TSP-1 and TGF-β expression was higher in the SCI group than in erythropoietin treatment group. Samples of spinal cord from the animals demonstrated a TSP-1 optical density of 112.2 ± 6.8 and TSP-1 positive cells were 5.7 ± 1.3 respectively. After injury, TSP-1 optical density and cell number increased to 287.2 ± 14.3 and 23.2 ± 2.6/mm2 at 24 h, 232.1 ± 13.2 and 15.2 ± 2.3/mm2 at 7 d. As to EPO treatment rats, the TSP-1 optical density and cell number decreased to 213.1 ± 11.6 and 11.9 ± 1.6/mm2 at 24 h, 189.9 ± 10.5 and 9.3 ± 1.5/mm2 at 7 d while comparing to the SCI rats (P < 0.01). TGF-β immunohistochemistry staining we couldn’t found it in sham-operated group rats. In SCI rats, optical density and positive neuron number were 291.4 ± 15.2 and 28.8 ± 4.9/mm2 at 24 h, 259.1 ± 12.3 and 23.9 ± 4.1/mm2 at 7 d, and also decreased in EPO treatment rats with 222.8 ± 11.9 and 13.7 ± 2.1/mm2 at 24 h, 196.5 ± 9.7 and 8.7 ± 2.2/mm2 at 7 d (P < 0.01). Conclusion Elevating expression of TSP-1 and TGF-β can be found in the injured segment of the spinal cord at 24h and 7d after injury, erythropoietin treatment effectively prevents pathological alterations from severer spinal cord injury via reduction the expression of TSP-1 and TGF-β.     

Hyperbaric oxygen intervention on expression of hypoxia inducible factor-1α and vascular endothelial growth factor after spinal cord injury models in rats

【Abstract】Objective: To observe the change of expression of Hypoxia inducible factor-1α (HIF-1α) and Vascular endothelial growth factor (VEGF) after spinal cord injury at different time and to investigate the Neuroprotective mechanism of hyperbaric oxygen (HBO) on spinal cord injury (SCI) in rats. Methods: 160 adult Sprague-Dawley rats, weighing between 250g and 300g,were randomly assigned to 4 experimental groups (n=40 per group). SCI group: SCI was created with special NYU impactor of Allen’s by a 25gram-centimeter (GCM) impacting energy on T10 of the spinal cord. SCI HBO group: hyperbaric oxygen therapy after spinal cord injury model was established. Sham operation（SH）group: only laminectomy of T10 and no impact on the spinal cord was done. SH HBO group: hyperbaric oxygen therapy after sham operation. The hindlimb functional recovery was evaluated using Basso Beattie Bresnahan (BBB) score and the expression of HIF-1α and VEGF were observed with fluorescent quantitation PCR and Western-Blot method of six rats picked randomly from each group at different times of 1, 3, 7 and 14 days after operation. Result: Rats in the SCI group and SCI HBO group were paralyzed completely after operation with BBB 0-1 score. Rats in the SH group and SH HBO group could walk after sham operation with BBB 20-21 score. The BBB scores of rats in SCI HBO group was higher than that in SCI group at 7d and 14d time point obviously (P＜0.05);The expression of HIF-1α and VEGF in SCI group and SCI HBO group was higher than in SH group and SH HBO group at any time point obviously (P＜0.05);while the SCI HBO group presented the least expression of HIF-1α at 3d,7d and 14d time points (P＜0.05) and more expression of VEGF at 7d and 14d (P＜0.05) than that of the SCI group with significant difference. Conclusion: The experimental research showed HBO could improve the hind limb functional recovery after SCI in rats;The higher expression of HIF-1α and VEGF could promote repair of damaged spinal cord after SCI;The elevation and duration of the expression of VEGF and the expression of HIF-1α by HBO intervention maybe inversely related in the repair of damaged spinal cord and neuroprotective effect.     

Correlation between magnetic resonance T2 image signal intensity ratio and cell apoptosis in a rabbit spinal cord cervical myelopathy model

Background Cervical spondylotic myelopathy （CSM） is a common cause of disability in elderly patients.Previous studies have shown that spinal cord cell apoptosis due to spinal cord compression plays an important role in the pathology of myelopathy.Although changes in magnetic resonance imaging （MRI） T2 signal intensity ratio （SIR） are considered to be an indicator of CSM,little information is published supporting the correlation between changes in MRI signal and pathological changes.This study aims to testify the correlation between MRI T2 SIR changes and cell apoptosis using a CSM animal model.Methods Forty-eight rabbits were randomly assigned to four groups：one control group and three experimental chronic compression groups,with each group containing 12 animals.Chronic compression of the cervical spinal cord was implemented in the experimental groups by implanting a screw in the C3 vertebra.The control group underwent sham surgery.Experimental groups were observed for 3,6,or 9 months after surgery.MRI T2-weighted SIR Tarlov motor scores and cortical somatosensory-evoked potentials （CSEPs） were periodically monitored.At each time point,rabbits from one group were sacrificed to determine the level of apoptosis by histology （n=6） and Western blotting （n=6）.Results Tarlov motor scores in the compression groups were lower at all time points than the control group scores,with the lowest score at 9 months （P ＜0.001）.Electrophysiological testing showed a significantly prolonged latency in CSEP in the compression groups compared with the control group.All rabbits in the compression groups showed higher MRI T2 SIR in the injury epicenter compared with controls,and higher SIR was also found at 9 months compared with 3 or 6 months.Histological analysis showed significant apoptosis in the spinal cord tissue in the compression groups,but not in the control group.There were significant differences in apoptosis degree over time （P ＜0.001）,with the 9-month group displaying the most severe spinal cord apoptosis.Spearman＇s rank correlation test showed that there was close relation between MRI SIR and degree of caspase-3 expression in Western blotting （r=0.824.P ＜0.001）.Conclusions Clear apoptosis of spinal cord tissue was observed during chronic focal spinal compression.Changes in MRI T2 SIR mav be related to the severity of the apoptosis in cervical spinal cord.     

首发抑郁症患者重复神经心理测查系统临床应用研究

Objective To investigate the damaging traits of cognitive function in first episode depression patients with the assessment of neuropsychological status ( RBANS) and Stroop Color-word Test. Methods Two hundred and thirty first-episode depression patients as experimental group and one hundred and seventy-eight health person as control were evaluated separately by repeatable battery for the RBANS and Stroop Color-word Test. Results In Stroop Color-word Test,the time required for completing the words,color,double-word,two-color in depression patients ( (14.37 ±6.65)s,(21. 58 ±8. 70) s,( 16. 56 ±8. 23)s, (37. 88 ± 13. 67) s) were longer than the control group ((12.38 ±3.34)s, (18.01 ±5.51) s, (14. 17 ±4. 37) s, (32. 87 ± 10.28) s). The difference between the two groups was found in cognitive function in first episode depression patients (P ＜ 0.05). The scores of color interference and meaning interference were no difference between the two groups (P＞0.05) . The patients'scores of the RBANS test in all items( (72.19 ± 17.22), (80.89 ± 15.29), (82.00 ± 16.37) , (98.74 ± 17.01) ,(82.09 ± 14.62) )were significantly poorer than those in healthy controls. Compared with control group, scores of the scales were significant difference between two groups. Pearson correlation analysis showed that RBANS total score and five factors had no correlation with HAMD total score and factors. It positively correlated with education level (P＜0.05) ,but not with sex (P＞0.05). Immediate memory,verbal function,delayed memory in five factors and RBANS total score negatively correlated with age and course of disease respectively, but breadth of vision,attention were not did(P＜0.05). Conclusion First-episode depression patients suffer from more serious and comprehensive damage of cognitive function such as memory,attention,executive function.     

Transplanted human umbilical cord blood mononuclear cells improve left ventricular function through angiogenesis in myocardial infarction

Background Human umbilical cord blood contains an abundance of immature stem/progenitor cells, which may participate in the repair of hearts that have been damaged by myocardial infarction (MI). This study aimed to evaluate the effects of human umbilical cord blood mononuclear cells (hUCBC) transplantation on cardiac function and left ventricular remodeling in rat model of MI. Methods Forty-five male Wistar rats were randomized into three groups: MI or control group (n=15), MI plus cell transplantation (n=15), and sham group (n=15). Acute myocardial infarction (AMI) was established by ligating the left anterior descending artery, thereafter, hUCBC were implanted into the marginal area of infarcted myocardium. In MI/control group, DMEM was injected instead of hUCBC following the same protocol. Left ventricular function assessment was carried out by echocardiography and invasive hemodynamic measurements one month post MI. All rats were sacrificed for histological and immunochemical examinations. Results The transplanted hUCBC survived and engaged in the process of myocardial repair in the host heart. Echocardiography demonstrated that left ventricular function improved significantly in the rats that underwent cell transplantation. Hemodynamic studies found a significantly decreased left ventricular end-diastolic pressure (LVEDP) [(21.08±8.10) mmHg vs (30.82±9.59) mmHg, P&lt;0.05], increase in +dp/dt(max) [(4.29±1.27) mmHg/ms vs (3.24±0.75) mmHg/ms, P&lt;0.05), and increase in －dp/dt(max) [(3.71±0.79) mmHg/ms vs (3.00±0.49) mmHg/ms, P&lt;0.05] among MI group with hUCBC transplantation when compared with MI/control group. Masson’s trichrome staining revealed that the collagen density in the left ventricle was significantly lower in rats of transplantation group than that in the MI control groups [(6.33±2.69)% vs (11.10±3.75)%, P&lt; 0.01]. Based on immunostaining of α-actin, the numbers of microvessels were significantly (P&lt;0.01) increased at the boundary of infarction site. Similarly higher mRNA expression of vascular endothelial growth factor (VEGF) 164 and VEGF188 were found at 7- and 28-day post cell transplantation in MI group with hUCBC transplantation when compared with MI/ control group.Conclusions Transplanted hUCBC can survive in host myocardium without immunorejection, significantly improve left ventricular remodeling after AMI and promote a higher level of angiogenesis in the infarct zones. All these factors beneficially affect cardiac repair in the setting of MI. Therefore human umbilical cord blood may be potential source for cell-based therapy for AMI.     

Human neural stem cells promote corticospinal axons regeneration and synapse reformation in injured spinal cord of rats

Background Axonal regeneration in lesioned mammalian central nervous system is abortive, and this causes permanent disabilities in individuals with spinal cord injuries. This paper studied the action of neural stem cell （NSC） in promoting corticospinal axons regeneration and synapse reformation in rats with injured spinal cord. Methods NSCs were isolated from the cortical tissue of spontaneous aborted human fetuses in accordance with the ethical request. The cells were discarded from the NSC culture to acquire NSC-conditioned medium. Sixty adult Wistar rats were randomly divided into four groups （n=15 in each）： NSC graft, NSC medium, graft control and medium control groups. Microsurgical transection of the spinal cord was performed in all the rats at the T11. The NSC graft group received stereotaxic injections of NSCs suspension into both the spinal cord stumps immediately after transection; graft control group received DMEM injection. In NSC medium group, NSC-conditioned medium was administered into the spinal cord every week; NSC culture medium was administered to the medium control group. Hindlimb motor function was assessed using the BBB Locomotor Rating Scale. Regeneration of biotin dextran amine （BDA） labeled corticospinal tract was assessed. Differentiation of NSCs and the expression of synaptophysin at the distal end of the injured spinal cord were observed under a confocal microscope. Group comparisons of behavioral data were analyzed with ANOVA. Results NSCs transplantation resulted in extensive growth of corticospinal axons and locomotor recovery in adult rats after complete spinal cord transection, the mean BBB scores reached 12.5 in NSC graft group and 2.5 in graft control group （P〈 0.05）. There was also significant difference in BBB score between the NSC medium （11.7） and medium control groups （3.7, P〈 0.05）. BDA traces regenerated fibers sprouted across the lesion site and entered the caudal part of the spinal cord. Synaptophysin expression colocalized with BDA positive axons and neurons distal to the injury site. Transplanted cells were found to migrate into the lesion, but not scatter along the route of axon grows. The cells differentiated into astrocytes or oligodendrocytes, but not into the neurons after transplantation. Furthermore, NSC medium administration did not limit the degree of axon sprouting and functional recovery of the injured rats compared to the NSC graft group. Conclusions Human embryonic neural stem cells can promote functional corticospinal axons regeneration and synapse reformation in the injured spinal cord of rats. The action is mainly through the nutritional effect of the stem cells on the spinal cord.     

Retraction: Prognostic analysis of transarterial chemoembolization combined with a traditional Chinese herbal medicine formula for treatment of unresectable hepatocellular carcinoma

Background  The most important objective of transplant studies in the injured spinal cord has been to provide a favorable environment for axonal growth. Moreover, the continuing discovery of new grafts is providing new potentially interesting transplant candidates. Our purpose was to observe the morphological and functional repair effects of the co-transplantation of neural stem cell (NSC), Schwann cells (SCs) and poly lactide-co-glycolide acid (PLGA) on the spinal cord injury of rats.

Methods  A scaffold of PLGA was fabricated. NSCs and SCs were cultured, with the NSCs labeled with 5-bromodeoxyuridine, and the complex of NSC/PLGA or NSC+SCs/PLGA were constructed. Thirty-six Wistar rats were randomly divided into three groups: group A (transplantation of PLGA), group B (transplantation of NSC/PLGA) and group C (transplantation of NSC+SCs/PLGA). The 3 mm length of the right hemicord was removed under the microscope in all rats. The PLGA or the complex of PLGA-cells were implanted into the injury site. Basso-Beattie-Bresnahan (BBB) locomotion scores, motor and somatosensory evoked potential of lower limbs were examined to learn the rehabilitation of sensory and motor function at 4 weeks, 8 weeks, 12 weeks and 24 weeks after injury. All the recovered spinal cord injury (SCI) tissues were observed with HE staining, immunohistochemistry, and transelectronmicroscopy to identify the survival, migration and differentiation of the transplanted cells and the regeneration of neural fibres at 4 weeks, 8 weeks, 12 weeks and 24 weeks after injury.

Results  (1) From 4 weeks to 24 weeks after injury, the BBB locomotion scores of cell-transplanted groups were better than those of the non-cell-transplanted group, especially group C (P <0.05). The amplitudes of the somatosensory evoked potential (SEP) and motor-evoked potential (MEP) were improved after injury in groups B and C, but the amplitude of SEP and MEP at 4 weeks was lower than that at 12 weeks and 24 weeks after injury. Compared with group B, the amplitude of SEP and MEP in group C was improved. The amplitude of SEP and MEP was not improved after injury in group A. (2) HE staining revealed the volume of the scaffold decreased and the number of cells in the scaffold increased. Newly-grown capillaries also could be seen. Immunohistochemistry staining showed the transplanted NSCs could survive and migrate until 24 weeks and they could differentiate into neurons and oligodendrocytes. The regenerated axons were observed in the scaffold-cell complex with transelectronmicroscopy. The above manifestations were more extensive in group C.

Conclusions  The transplanted NSC can survive and migrate in the spinal cord of rats up to 24 weeks after injury, and they can differentiate into various neural cells. Co-transplantation of cells/PLGA can promote the functional recovery of the injured spinal cord. The effect of co-transplanting NSC+SCs/PLGA is better than transplanting NSC/PLGA alone.

     

年龄对嗅鞘细胞移植治疗脊髓损伤疗效的影响

为探讨年龄因素对嗅鞘细胞移植治疗脊髓损伤临床疗效的影响 ,用嗅鞘细胞 (OECs)移植到脊髓损伤部位上下处的方法 ,共治疗 1 71例脊髓损伤 0 .5～ 1 8年的患者。患者年龄 2 .5～ 64岁 (平均 3 4.9岁 ) ,按年龄分为 5组 :≤ 2 0岁 9例 ,2 1～ 3 0岁 5 4例 ,3 1～ 40岁 60例 ,41～ 5 0岁 3 4例 ,≥ 5 1岁 1 4例。嗅鞘细胞移植后 2～ 8周随访 ,根据美国脊髓损伤学会 (ASIA)脊髓损伤神经功能分类国际标准评价。结果 :1 71例患者的脊髓功能均有部分恢复 ,除 5 1岁以上组的痛觉恢复与其他年龄组 (除 2 1～ 3 0岁组 )比较有显著性差异 (P <0 .0 5 )外 ,各年龄组的运动和触觉改善比较均无显著性差异。结果提示 :嗅鞘细胞移植能改善所有年龄段脊髓晚期损伤患者的神经功能 ,除老年组痛觉恢复较好外 ,年龄因素对运动和触觉的近期效果无明显影响 ,长期结果有待进一步研究     

嗅鞘细胞抑制脊髓损伤后硫酸软骨素蛋白聚糖表达的实验研究

目的:探讨嗅鞘细胞(OECs)移植的治疗作用及其对影响硫酸软骨素蛋白聚糖类分子(CSPGs)表达的影响。方法:将培养的OECs移植于脊髓半横断的Wistar大鼠,移植术后对模型进行数个时间点的BBB运动功能评分,损伤即移植术后4周,对实验组和对照组动物取材,进行免疫组织化学和Western blotting检测。结果:移植后4周,实验组动物的BBB评分高于对照组动物。免疫组织化学和Western blotting检测发现,实验组动物损伤处CSPGs物质表达低于对照组动物。结论:OECs促进脊髓损伤动物运动功能恢复的作用可能与抑制CSPGs分子表达有关。     

肋间神经移植加NGF、BDNF基因修饰的OECs修复大鼠脊髓损伤

目的探讨自体肋间神经移植加NGF、BDNF基因修饰的嗅神经鞘细胞（OECs）修复大鼠脊髓损伤（Spinal cord injury，SCI）。方法SD大鼠72只，经T11、T12间隙平面切取2mm的半侧脊髓，制成半切伤模型。动物分为A、B、C三组，每组24只。A组：将自体肋间神经纤维损伤脊髓近端前角灰质与远端前索白质桥接连接，伤侧L1、L2、L3、L4脊神经节与近端后索白质及远端后角灰质与近端后索白质桥接连接，并植入NGF、BDNF基因修饰的OECs；B组：同上述方法移植肋间神经，不植入OECs；C组：单纯脊髓损伤，脊髓半切缺损处填充明胶海棉，术后4、8周对各组动物进行神经功能检查（斜板实验检查，BBB评分），感觉诱发电位（SEP）和运动诱发电位（MEP）检测以及组织学检查。结果术后4周时，A、B组和动物神经功能，斜板试验，BBB评分及SFP、MEP检测结果均优于C组，8周时，A组动物神经功能，斜板试验，BBB评分，SEP、MEP检测结果均优于B组和C组，镜下见移植组织存活，充填于缺损处，部分脊髓组织融合，结论肋间神经移植加NGF、BDNF基因修饰的OECs对SCI的修复有一定的疗效。     

嗅鞘细胞移植联合应用GDNF对大鼠脊髓损伤的治疗作用

目的：研究嗅鞘细胞（OECs)移植并联合应用胶质细胞源性神经营养因了（GDNF）对脊髓损伤的保护和促进再生作用。方法；建立成年SD大鼠脊髓T8半横断损伤模型，将体外培养纯化的OECs植入脊髓损伤处，同时局部应用GDNF，采用斜板试验和BBB联合功能评分观察大鼠运动功能恢复情况，并通过辣根过氧化物酶逆行示踪技术评价OECs和GDNF对神经元存活和纤维再生的影响。结果：（1）OECs和GDNF联合应用时对皮质和红核神经元有逆行性保护作用；可促进脊髓下行传导束再生，肢体运动功能恢复，（2）OECs和GDNF联合应用组对脊髓损伤的修复作用最强，高于GDNF或OECs单用组。结论：联合应用GDNF和OECs在脊髓损伤修复治疗中具有协同作用。     

嗅鞘细胞复合细胞外基质凝胶移植对损伤脊髓细胞的保护作用

目的:探讨嗅鞘细胞(OECs)和细胞外基质(ECM)凝胶复合移植对损伤脊髓细胞的保护作用。方法:将脊髓半横断伤SD大鼠模型,随机分为嗅鞘细胞移植组(A组)、损伤对照组(B组)和正常组(C组),24h每组8只动物取伤段标本测水离子含量。其余动物第6周、第12周每组8只动物爬坡试验,评价下肢运动功能及运动诱发电位(MEP)检测。结果:脊髓损伤(SCI)后组织水肿,Na+、Ca2+离子浓度升高,K+、Mg2+离子浓度降低。嗅鞘细胞复合细胞外基质凝胶脊髓内移植后显著改善这些变化,使损伤细胞功能有显著恢复。结论:嗅鞘细胞复合细胞外基质凝胶脊髓内移植对SCI有保护作用,其机制可能与改善细胞内环境有关。     

嗅神经鞘细胞移植治疗实验性脊髓损伤的研究

目的:观察移植体外培养的嗅神经鞘细胞(OEC)对大鼠脊髓左半侧横断伤的治疗作用。方法:将应用改良Nash法培养的OECs移植到成年大鼠T10脊髓左半侧横断处,8周后分别采用改良Tarlov法运动学功能评分、辣根过氧化物酶(HRP)顺行示踪、透射电镜观察、P75免疫组织化学染色及HE染色等方法,观察动物的运动功能、脊髓再生的轴突和髓鞘、OEC的存活和脊髓损伤程度。结果:8周时治疗组运动学功能评分高于对照组(P<0.05);治疗组再生轴突穿越损伤区,对照组则无再生轴突穿越;超微结构显示治疗组轴突数量多于对照组,且髓鞘的完整性强于对照组,OEC表现为星形胶质细胞样和雪旺氏细胞样两种,包绕轴突髓鞘;免疫组化染色显示移植后8周OECs仍然存活,在损伤区均匀、散在分布;损伤后8周脊髓损伤区有胶质瘢痕及空洞形成。结论:移植的嗅神经鞘细胞可促进大鼠运动功能的恢复、脊髓轴突的再生和髓鞘的重塑。     

嗅鞘细胞移植后脊髓挫伤大鼠几种神经营养因子的表达变化

目的探讨嗅鞘细胞移植脊髓挫伤大鼠后,在其神经损伤及修复过程中三种神经营养因子（脑源性神经营养因子、神经生长因子、睫状神经营养因子）的表达变化及意义。方法体外分离培养大鼠嗅球嗅鞘细胞。40只健康SD大鼠,雌雄不拘,在移植前一星期使用NYU-撞击机,利用10g-25mm的致伤力,制作T10脊髓挫伤模型。40只脊髓挫伤大鼠随机分为OECs移植组和对照组（注射无血清培养液组）,又根据取材时间不同分为术后1天、7天、14天、21天组。动物处死前进行BBB评分评估后肢运动功能的变化。取手术部位头端和尾端0.5cm脊髓,采用免疫组织细胞化学染色检测技术检测三种神经营养因子（BDNF、CNTF、NGF）在各组脊髓中的表达变化。结果行为学评分随着损伤时间的推移呈小幅度增加,但后肢无运动功能恢复;免疫组织化学检测显示,嗅鞘细胞移植能部分改善大鼠脊髓灰质神经细胞的凋亡程度,延缓白质神经纤维的减少,促进髓鞘的修复与再生。与对照组比较,嗅鞘细胞移植治疗后脊髓组织中三种神经营养因子表达明显增加（P〈0.05）。结论嗅鞘移植能够部分改善脊髓损伤后脊髓组织的病理形态,促进脊髓组织中多种神经营养因子的表达。     

NTFs基因修饰的人胚OECs植入促进大鼠SCI神经再生及功能恢复的研究

目的探讨神经营养因子（NTF）基因修饰嗅鞘细胞（OECs）移植促进大鼠脊髓损伤后神经再生及功能恢复。方法SD大鼠制备成脊髓损伤动物模型。随机分为损伤对照组（A组）、OECs移植组（B组）和OECs神经营养因子基因修饰后移植组（C组）。治疗后2、4、6周,每组动物分别进行联合行为评分（GBS）、运动诱发电位（MEP）、感觉诱发电位（SEP）检查、双下肢功能测定（爬坡试验）,评价脊髓损伤功能恢复情况。结果随时间的延长,B组GBS、MEP、SEP及下肢功能明显改善。与其它组相比较,差异有显著性,P〈0.05。结论NTF基因修饰后移植在脊髓损伤区,能促进神经再生及功能恢复。     

BMSCs并BDNF移植治疗大鼠陈旧性脊髓损伤效果

目的探讨骨髓间充质干细胞(BMSCs)并脑源性神经营养因子(BDNF)移植治疗大鼠陈旧性脊髓损伤的效果。方法对48只Wistar大鼠采用改良的ALLEN撞击法建立脊髓损伤模型,将其随机分为4组,4周后4组分别注射BMSCs+BDNF、BMSCs、BDNF及DMEM干预。分别于移植后1、2、4周采用BBB评分评估大鼠后肢运动情况,并于移植后4周采用免疫组化法检测BMSCs在脊髓损伤部位的分布,及生长相关蛋白43(GAP-43)的表达。结果移植后4周,BMSCs+BDNF组及BMSCs组的BMSCs在脊髓损伤部位均得以存活。BMSCs+BDNF组与BMSCs组及BDNF组比较,脊髓空洞较少并且GAP-43阳性表达面积较高(F=35.57,q=4.97～25.84,P<0.05)。BMSCs+BDNF组BBB评分与其他组比较,差异有显著性(F=27.51,q=3.89～11.57,P<0.05)。结论 BMSCs+BDNF联合移植对于大鼠陈旧性脊髓损伤有较好的修复作用。     

活化巨噬细胞移植时间和脊髓损伤后功能恢复的实验研究

目的：探讨脊髓损伤后活化巨噬细胞移植的时机和脊髓功能恢复之间的关系。方法：将62只脊髓左侧半切的BALB C小鼠随机分为A、B、C三组，A、B组小鼠各21只，分别在脊髓半切后1h和8h移植活化的巨噬细胞。C组20只小鼠，为对照组。随机各取8只小鼠，在脊髓半切后的每周观察其左后肢运动功能并评分（BBB）、右后肢感觉诱发电位（SEP）的潜伏期和波幅数值的变化情况。此外，随机从每组抽取8只小鼠，分别在第1天、第4天、第7天、第2周、第4周、第8周、第12周、第16周处死后行常规HE染色和免疫组化染色，观察巨噬细胞、胶质细胞和神经元轴突数量的变化。结果：A组小鼠和B、C组小鼠运动功能评分以及SEP潜伏期和波幅数值经单因素的方差检验差异有显著性（P〈O．05）。A组小鼠神经元细胞染色阳性级别明显增高（P〈0．05）。结论：采用一定浓度的巨噬细胞移植促进损伤脊髓功能的恢复．需要合适的时机．